Development of animal model enabling the manipulation of intracellular Ca level

为了实现细胞内Ca2+水平的细胞型特异性操纵，我们建立了黑视素（人类OPN4基因）以细胞型特异性表达的转基因动物。我们培育了tetO-melanopsin转基因小鼠，并与神经元tTA （orexin-tTA）小鼠杂交。食欲素神经元表达了足够数量的黑视素，从而引起细胞放电，证明了成功的操作。与神经胶质小鼠杂交时，无诱导作用。为了改善黑视素在神经胶质细胞中的诱导作用，我们利用小鼠黑视素位点产生STOP tetO-melanopsin敲入小鼠。与之前敲入tetO的小鼠不同，还没有神经胶质细胞系诱导黑视素。我们能够在神经元细胞中表达足够数量的黑视素，并通过光照操纵细胞功能，但我们无法在胶质细胞中表达黑视素。

To achieve the manipulation of intracellular Ca2+ levels in the cell-type specific manner, we established the transgenic animal in which melanopsin (human OPN4 gene) was expressed in cell-type specific manner. We generated tetO-melanopsin transgenic mice and crossed with neuronal tTA (orexin-tTA) mice. Orexin neurons expressed enough amount of melanopsin to evoke cell firing, demonstrating the successful manipulation. In case of crossing with glial tTA mice, there was no induction. To improve melanopsin induction in glial cells, we generated STOP tetO-melanopsin knockin mice using mouse melanopsin locus. Unlike previous tetO knockin mice, none of glial tTA line induced melanopsin yet. We were able to express sufficient amount of melanopsin in neuronal cells and manipulate cell function by illumination, however, we failed to express melanopsin in glial cells.